phage - ubc - nls - mcp - egfp - BirA*
(Plasmid #131134)

• Purpose: BirA* expressed as a protein fusion with MS2 coat protein (MCP) monomer and EGFP; to visualize any MS2 tagged mRNAs and to identify the RNA proximal proteome by their biotinylation.
• Depositing Lab: Ralf Jansen
• Publication: Jansen lab plasmids (unpublished)

Backbone

• Vector backbone: pHAGE-UBC
• Backbone size w/o insert (bp): 6248
• Vector type: Mammalian Expression, Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: NLS-HA-MCP-EGFP-BirA*
• Alt name: NLS-HA-MCP-EGFP-BioID
• Species: Synthetic
• Insert Size (bp): 2493
• Mutation: BirA* = BIrA >R118G
• Promoter: human ubiquitin C (UBC)
• Tag / Fusion Protein:
  • NLS, HA (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: ClaI (not destroyed)
• 5′ sequencing primer: 5'-ATTACAGGGACAGCAGAGATCC-3'
• 3′ sequencing primer: WPRE-R (5'CATAGCGTAAAAGGAGCAACA-3')

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Variant of phage - ubc - nls - 2xmcp - egfp - BirA* (Addgene plasmid 131132 ) with one copy of MCP protein compared to two MCPs. Consequently, binding of two MCPs per MS2 RNA loop results in twice the number of BirA* proteins than in previous cassettes